Epidemiology and molecular evolution of Paslahepevirus balayani (Hepatitis E Virus) in fur-bearing animals in Hebei Province, China

Hepatitis E virus (HEV) is a small, non-enveloped RNA virus primarily transmitted via the fecal-oral route, causing large-scale outbreaks or sporadic infections. With the increasing farming and trade of fur-bearing animals in China, potential zoonotic risks have gained attention. This study aimed to investigate the prevalence and epidemiological trends of HEV infection in farmed fur-bearing animals (raccoon dogs, mink, and foxes) in Hebei Province, China. A total of 272 liver and fecal samples were collected from fur farms. HEV RNA was detected using RT-qPCR and nested RT-PCR, followed by amplification of the ORF2 region and whole-genome sequencing. Infection rates were calculated, and phylogenetic analysis was performed on the obtained sequences. Overall, 43 samples (15.81%) tested positive for HEV RNA. The infection rates were 19.35% (36/186) in raccoon dogs and 8.64% (7/81) in mink; no HEV RNA was detected in foxes. A total of 24 ORF2 fragments and 11 full-genome sequences were obtained. Phylogenetic analysis showed that all full-genome sequences belonged to HEV genotype 4 and formed two distinct branches. One branch clustered with HEV-4d and was closely related to swine-derived HEV sequences from Gansu Province, China. The other branch exhibited significant genetic divergence from known HEV-4 subtypes, potentially representing a new subtype or variant. This study systematically investigated HEV prevalence in fur-bearing animals and, for the first time, obtained the complete genome sequence of an HEV-4 isolate from mink in Hebei Province. This indicates that there may be a new subtype of HEV-4 in raccoon dogs, which provides the latest data of HEV infection and circulation in fur animals in Hebei Province and supports the formulation of effective public health policies.